rhGCSF對小鼠骨髓間充質(zhì)干細(xì)胞的動(dòng)員作用.doc

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1、rhGCSF對小鼠骨髓間充質(zhì)干細(xì)胞的動(dòng)員作用作者：劉岐煥，程范軍，陳龍，唐俊明，王家寧，高清平【摘要】為了研究rhGCSF體內(nèi)應(yīng)用對小鼠骨髓間充質(zhì)干細(xì)胞（MSC）的動(dòng)員作用，將30只小鼠隨機(jī)分為2組：rhGCSF組和對照組，分別給予皮下注射rhGCSF80μg/（kg·d）和等量生理鹽水,連續(xù)5天，在最后1次注射后分別于6，12和168小時(shí)取小鼠骨髓和外周血，以1×106單個(gè)核細(xì)胞（MNC）接種于12孔培養(yǎng)板14天，對形成的成纖維細(xì)胞集落（CFUF）進(jìn)行計(jì)數(shù)，用胰蛋白酶消化后應(yīng)用流式細(xì)胞術(shù)測定表面抗原并進(jìn)行成脂，成骨誘導(dǎo)。結(jié)果表明：rhGCSF組骨髓形成的CFUF數(shù)

2、均較對照組明顯增加(p<0.01)。6小時(shí)取材較12小時(shí),168小時(shí)取材形成的CFUF數(shù)目多(p<0.01)，12小時(shí)與168小時(shí)CFUF差異無統(tǒng)計(jì)學(xué)意義。流式細(xì)胞術(shù)檢測骨髓CFUF顯示CD34－、CD133－、CD90＋、CD105＋，骨髓CFUF具有成脂、成骨分化的能力。rhGCSF組6小時(shí)取材外周血具有類似骨髓的CFUF，出現(xiàn)的頻率為（0.50±0.11）×10-6，與對照組比較差異有顯著性（p<0.05）。結(jié)論：rhGCSF可使小鼠骨髓間充質(zhì)干細(xì)胞增殖加速，但時(shí)間短暫，其高峰在rhGCSF應(yīng)用5天后的6小時(shí)內(nèi)。rhGCSF對小鼠間充質(zhì)干

3、細(xì)胞也有動(dòng)員能力，但作用較弱?！娟P(guān)鍵詞】骨髓間充質(zhì)干細(xì)胞EffectsofrhGCSFonMobilizationofMouseMesenchymalStemCellsAbstractToevaluatetheeffectsofrhGCSFonmobilizationofmesenchymalstemcells(MSCs)ofmousebonemarrowatdifferenttimepoint,thirtymicewererandomlydividedintorhGCSFtreatmentgroupandcontrolgroup.Themiceweresubcutaneo

4、uslyinjectedwithrhGCSFinadoseof80μg/kgorsalinefor5days.Thebonemarrowandperipheralbloodwereobtainedattimepointsof6,12,168hoursafterfinalinjectionofrhGCSForsaline.Bonemarrowmononuclearcells(BMMNCs)wereseededatdensityof1×106MNCsonto12wellplateforcultureexpansioninDMEMsupplementedwith10%FBS,an

5、dthenumberofcolonyformingunitfibroblast(CFUF)wascountedafter14days.ThecellswerecollectedbytrypsinizationandthesurfaceantigensCD34,CD133,CD90andCD105wereanalyzedbyflowcytometry.ThemultidifferentiationofMSCsweredoneinthecultureconditionofinducedadipocyteandosteocyte.PeripheralbloodMNCsexami

6、nationwassameasthebonemarrow.TheresultsindicatedthatthenumberofCFUFofbonemarrowinrhGCSFgroupwasmorethanthatincontrolgroup(p<0.01)，thenumberofCFUFinrhGCSFgroupat6hourswasmorethanthatat12hoursand168hours,7respectively(p<0.01).TherewasnoobviousdifferencebetweenCFUFat12hoursandat168hou

7、rs(p>0.05).MSCswerepositiveforCD90,CD105andnegativeforCD34andCD133.MSCswerefoundtodifferentiateintoadipocyteandosteocyteinvitro.TheCFUFofPBMNCsobtainedandculturedinvitrointhesamecultureconditionscouldbeobservedaftertherhGCSFinjectionat6h